Nanovesicles derived from bacteria of lactococcus genus and use thereof
A technology of lactococcus and vesicles, applied in streptococcus/lactococcus, bacteria used in food preparation, medical preparations containing active ingredients, etc.
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Embodiment 1
[0100] Example 1. Analysis of in vivo absorption, distribution and excretion patterns of bacteria and vesicles derived from bacteria
[0101] To assess whether gut bacteria and bacterial-derived vesicles are absorbed systemically through the gastrointestinal tract, the following experiments were performed. First, a dose of 50 μg of fluorescently labeled enterobacteria and enterobacteria-derived vesicles were each administered to the stomach of mice via the gastrointestinal tract, and fluorescence was measured after 0 min, 5 min, 3 h, 6 h, and 12 h. As a result of observing the whole image of the mouse, as Figure 1A As shown, bacteria were not taken up systemically, but vesicles derived from bacteria were absorbed systemically at 5 minutes after administration, and strong fluorescence was observed in the bladder at 3 hours after administration, making it possible to see that the vesicles were excreted. to the urinary tract. Furthermore, it can be seen that vesicles are pres...
Embodiment 2
[0103] Example 2. Metagenomic Analysis of Bacteria-Derived Vesicles in Clinical Samples
[0104] After first putting the clinical sample (such as blood, urine or saliva, etc.) test tube. After removing bacteria and impurities using a 0.22-μm filter, transfer them to a Centriprep tube (centrifugal filter 50kD) and centrifuge at 1,500×g and 4°C for 15 minutes, discard the material smaller than 50kD, and the residue Concentrate to 10ml. After bacteria and impurities were removed again using a 0.22-μm filter, supernatant was discarded and aggregated pellets were dissolved by performing ultracentrifugation at 150,000×g and 4°C for 3 hours using a 90Ti type rotor in normal saline (PBS).
[0105] Internal DNA was extracted from lipids by boiling 100 μl of vesicles isolated by the method described above at 100 °C, followed by cooling on ice for 5 min. Then, in order to remove the residual suspension, the DNA was centrifuged at 10,000×g and 4°C for 30 minutes, and only the supern...
Embodiment 3
[0109] Example 3. Metagenomic Composition of Bacteria and Bacteria-Derived Vesicles in Saliva of Diabetics and Normal Individuals analysis
[0110] After metagenomic analysis of saliva from 47 diabetic patients and 277 age- and sex-matched normal individuals by extracting genes from bacteria and vesicles present in saliva using the method of Example 2, milk Distribution of Bacillus and Lactococcus-derived vesicles. As a result, it was confirmed that bacteria of the genus Lactococcus and vesicles derived from bacteria of the genus Lactococcus were significantly reduced in the saliva of diabetic patients compared with the saliva of normal individuals (see Tables 2, 3 and figure 2 ).
[0111] _[Table 2]
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[0113] [table 3]
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